(A) is the schematic of morphine dependent zebrafish CPP experiment; (B) is the schematic of morphine dependent zebrafish CPP experiment with antibiotic treatment.

Time spent by zebrafish in the drug-pair compartment. (n = 8). ^∗∗P < 0.01 vs the C group; ^##P < 0.01 vs the M group. C, control group; M, morphine group; M+S, morphine+sinomenine group.

Effect of sinomenine use on the gut microbiome accompanying morphine dependence. (A) Venn diagram depicting the distribution of bacterial taxa (at the OTU level) from the different zebrafish experimental groups. (B) Alpha diversity of the fecal microbiome among the three groups according to the Chao 1, Richness, Shannon, and Simpson indices. (C) Beta diversity of the fecal microbiome among the three groups based on principal coordinate analysis (PCoA) and nonmetric multidimensional scaling (NMDS) at the OTU level based on Bray–Curtis similarity algorithms. (D) Comparison of species composition among the fecal microbiota of the C, M, and M+S groups at the phylum level. (E) Prediction of changed KEGG pathways using PICRUST analysis. *P < 0.05, ^∗∗P < 0.01 vs the C group; ^#P < 0.05, ^##P < 0.01 vs the M group. C, control group; M, morphine group; M+S, morphine+sinomenine group.

qPCR analysis of occludin a, occludin b, il1b, oprm1, oprd1, drd2a, htr2aa, bdnf, and ntrk2 expression in zebrafish. (n = 3). ^∗P < 0.05, ^∗∗P < 0.01 vs the C group; ^#P < 0.05, ^##P < 0.01 vs the M group. C, control group; M, morphine group; M+S, morphine+sinomenine group.

Time spent by zebrafish in the drug-pair compartment. (n = 8). ^∗∗P < 0.01 vs the A group. A, antibiotic group; A+M, antibiotic+morphine group; A+M+S, antibiotic+morphine+sinomenine group.

Effect of antibiotic use on the sinomenine treatment of the morphine-dependent gut microbiome. (A) Venn diagram depicting the distribution of bacterial taxa (at the OTU level) in the different experimental groups of zebrafish. (B) Alpha diversity of the fecal microbiome among the three groups according to the Chao 1, Richness, Shannon, and Simpson indices. (C) Beta diversity of the fecal microbiome among the three groups based on principal coordinate analysis (PCoA) and nonmetric multidimensional scaling (NMDS) at the OTU level based on the Bray–Curtis similarity algorithms. (D) Comparison of species composition among the fecal microbiota of the A, A+M, and A+M+S groups at the phylum level. (E) Prediction of changes in KEGG pathways using PICRUST analysis. *P < 0.05, **P < 0.01 vs the A group; ^#P < 0.05, ^##P < 0.01 vs the A+M group. A, antibiotic group; A+M, antibiotic+morphine group; A+M+S, antibiotic+morphine+sinomenine group.

qPCR analysis of occludin a, occludin b, il1b, oprm1, oprd1, drd2a, htr2aa, bdnf, and ntrk2 expression in zebrafish. (n = 3). *P < 0.05, **P < 0.01 vs the A group; ^#P < 0.05, ^##P < 0.01 vs the A+M group. A, antibiotic group; A+M, antibiotic+morphine group; A+M+S, antibiotic+morphine+sinomenine group.